Fine mapping and discovery of candidate genes underlying seed size are important for modern watermelon breeding. Here, by using a high-resolution genetic map and whole-genome genetic variation detection aided by genome survey sequencing, we fine mapped and discovered candidate genes for seed size in watermelon. QTL (quantitative trait locus) mapping identified two pleiotropic QTLs for seed size, namely, qSS4 and qSS6, using a high-density genetic map constructed by specific length amplified fragment sequencing. qSS6 explained 93.00%, 94.11% and 95.26% of the phenotypic variation in thousand-seed weight, seed length and seed width, respectively, and was defined as a major QTL. Then, high-coverage re-sequencing of two parental lines detected a total of 193,395 SNPs (single nucleotide polymorphisms) and 45,065 indels (insertions/deletions), which corresponded to a frequency of 534 SNPs/Mb and 124 indels/Mb. Based on the genetic variation in the two parental lines, newly developed PCR-based markers allowed the region of qSS6 to be narrowed to 55.5 kb. Three potential candidates were identified, including a known seed size regulator in rice, SRS3. Taken together, our results reveal successful rapid fine mapping and discovery of candidate genes for seed size in watermelon, which could be applied to many traits of interest in plants.